Method for rapidly and comprehensively detecting oligosaccharide on medicinal monoclonal antibody N-glycosylation site

Abstract

The invention provides a method for rapidly and comprehensively detecting oligosaccharide on a medicinal monoclonal antibody N-glycosylation site. The method comprises the following steps: (1) sample oligosaccharide enzymolysis; (2) 2-AB derivatization solution preparation; (3) derivatization of the oligosaccharide; (4) purification of derived oligosaccharide; and (5) LC-fluorescence-ESI-MS analysis of the derived oligosaccharide. According to the method provided by the invention, monoclonal antibody N-glycosylation can be effectively, rapidly and accurately analyzed; and analysis and production quality control on the medicinal monoclonal antibody molecule action mechanism are facilitated.

Description

technical field [0001] The invention belongs to the field of biological genetic engineering, and in particular relates to a method for detecting oligosaccharides on the N-glycosylation site of a medicinal monoclonal antibody. Background technique [0002] N-glycosylation on a pharmaceutical monoclonal antibody (mAb) molecule has an important impact on its stability and activity. Detection of oligosaccharides at N-glycosylation sites is also critical for monitoring batch stability of pharmaceutical mAb production. The currently used detection method for oligosaccharides on the glycosylation site of monoclonal antibody is: use a specific enzymatic reaction (such as using peptide N glycosidase PNGase f) to remove the oligosaccharides on the glycosylation site, and use solid phase extraction and other methods for purification, and subsequent detection includes: (1) Fluorescently label the purified oligosaccharides (such as 2-AA, 2-AB and other fluorescent reagents), and use HIL...

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Problems solved by technology

It is not possible to deduce oligosacch

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